抑制泛素羧基末端水解酶L1 对小鼠脑缺血/再灌注损伤的影响

doi:10.16098/j.issn.0529-1356.2021.04.007

摘要/Abstract

摘要：

目的评估抑制泛素羧基末端水解酶L1(UCHL1)对小鼠脑缺血/再灌注损伤的影响。方法将雄性BALB/c小鼠随机分为假手术（sham）组、缺血/再灌注(I/R)组、UCHL1小干扰 RNA (siRNA) 组和scramble siRNA(control) 组，每组10只小鼠。采用小鼠大脑中动脉闭塞(MCAO) 60 min后再灌注24 h建立I/R模型。其中siRNA组和control组在MCAO前24 h将10 μl UCHL1 siRNA或scramble siRNA通过侧脑室注入脑内。采用RT-PCR和Western blotting方法检测各组小鼠脑组织中UCHL1表达；采用2，3，5-氯化三苯基四氮唑（TTC）染色法评估各组小鼠脑梗死体积和水肿率；采用神经行为学评分法评估各组小鼠神经症状评分。结果与sham组相比，I/R组缺血半影区UCHL1 mRNA和蛋白水平显著增高(P<0.05)；而 siRNA组UCHL1蛋白和mRNA表达明显降低(P<0.05)；与此同时，I/R组小鼠脑梗死体积、水肿率及神经行为学损伤明显增加，而siRNA组小鼠脑梗死体积和水肿率及神经行为学损伤进一步加重(P<0.05)。结论抑制UCHL1可加重小鼠脑缺血/再灌注损伤。提示，MCAO后诱导UCHL1表达对小鼠脑缺血/再灌注损伤具有保护作用。

关键词: 泛素羧基末端水解酶L1, 缺血/再灌注损伤, 反转录聚合酶链反应, 免疫印迹法, 神经行为学评分, 2,3,5-氯化三苯基四氮唑染色, 小鼠

Abstract:

Objective To evaluate the effect of inhibition of ubiquitin carboxy terminal hydrolase L1 (UCHL1) on cerebral ischemia/reperfusion injury in mice. Methods Male BALB/c mice were randomly divided into sham group, ischemia/reperfusion (I/R) group, UCHL1 small interfering RNA (siRNA）group and scramble siRNA (control) group, 10 mice in each group. I/R model was established by reperfusion 24 hours after middle cerebral artery occlusion (MCAO）60 minutes. In the siRNA group and control group, 10 μl UCHL1 siRNA or scramble siRNA was injected into the brain through the lateral ventricle 24 hours before MCAO. The expression of UCHL1 was detected by RT-PCR and Western blotting; the volume of cerebral infarction and the rate of edema were assessed by 2,3,5-triphenyl tetrazolium chloride (TTC) staining; and the score of neurological symptoms was assessed by neurobehavioral scoring. Results Compared with the sham group, the level of UCHL1 mRNA and protein in ischemic penumbra of I/R group were significantly higher (P<0.05), while the expression of UCHL1 protein and mRNA in siRNA group were significantly lower (P<0.05); at the same time, the volume of cerebral infarction, edema rate and neurobehavioral damage in I/R group increased significantly, while the volume and edema rate of cerebral infarction and neurobehavioral damage in siRNA group further increased (P<0.05). Conclusion Inhibition of UCHL1 can aggravate the cerebral ischemia/reperfusion injury in mice, suggesting that the induction of UCHL1 after MCAO has a protective effect on the cerebral ischemia/reperfusion injury in mice.

Key words: Ubiquitin carboxy terminal hydrolase L1, Ischemia/reperfusion injury, RT-PCR, Western blotting, Neurobehavioral score, 2,3,5-triphenyl tetrazolium chloride staining, Mouse

中图分类号:

R363.1

彭志锋李晨旭马国英杨靖辉. 抑制泛素羧基末端水解酶L1 对小鼠脑缺血/再灌注损伤的影响[J]. 解剖学报, 2021, 52(4): 543-547.

PENG Zhi-feng LI Chen-xu MA Guo-ying YANG Jing-hui. Inhibition effect of ubiquitin carboxy terminal hydrolase L1 on cerebral ischemia/reperfusion injury in mice[J]. Acta Anatomica Sinica, 2021, 52(4): 543-547.

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